1. ¹Department of Dermatology, Eberhard Karls University, Tuebingen, Germany
2. ²Department of Dermatology, Ludwig Maximilians University, Munich, Germany
3. ³Department of Dermatology, AK St Georg, Hamburg, Germany

Correspondence: Dr Christian A. Sander, Department of Dermatology, AK St Georg, Lohmuehlenstr. 5, 20099 Hamburg, Germany. E-mail: Christian.Sander@ak-stgeorg.lbk-hh.de

Received 11 April 2005; Revised 19 August 2005; Accepted 26 August 2005; Published online 22 December 2005.

Abstract

It is established that primary malignant melanomas (pMM) can be infiltrated by T-cell populations with predominantly one T-cell clone. As pMM generally express multiple tumor-associated antigens (TAA), here we used laser-capture microdissection (LCM) to isolate different tumor-infiltrating lymphocyte (TIL) clusters in order to determine whether pMM are infiltrated only by one single clone or whether the TAA may attract various T-cell populations. As T-cell receptor (TCR) clonality is a useful tool for the demonstration of specific T-cell clones, we analyzed 56 pMM, three cutaneous melanoma metastases, and 15 pairs of pMM with a sentinel lymph node (SLN) for clonal rearrangements of the (TCR) chain gene. We detected the clonality of TCR chain gene in 25 of 56 pMM, and in 10 of 17 SLN studied. In four of the 15 pairs of primary tumor and SLN, we found clonal TCR in both the melanoma and the SLN, with two pairs harboring the identical clone. As we detected different clones in pMM and the corresponding SLN, we subsequently performed LCM in 21 malignant melanomas with multiple lymphocytic clusters for the presence of focal clonal T cells in different regions of the melanoma. In seven melanomas, both clusters of TILs showed the same rearranged TCR chain gene and in five of the seven biopsies the clonal rearrangement occurred in different variable (V) regions of the TCR chain gene. These tumors showed infiltration by more than one clone. In 10 biopsies TCR clonality was restricted to one cluster, while the second microdissected sample of the infiltrate was polyclonal. In conclusion, within one primary malignant melanoma several T-cell clones with different rearrangements may occur. The balance between these clones may decide on the progress of melanoma.